| Cav2.2e37a基因RNAi慢病毒载体的构建与鉴定 |
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| 引用本文： | 王维,冯泽国,陈娜,马涛,周建平.Cav2.2e37a基因RNAi慢病毒载体的构建与鉴定[J].军医进修学院学报,2010,31(10):1016-1018. |
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| 作者姓名： | 王维 冯泽国 陈娜 马涛 周建平 |
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| 作者单位： | [1]解放军总医院麻醉手术中心,北京100853 [2]第二炮兵总医院麻醉科,北京100088 [3]军事医学科学院毒物药物研究所,北京100850 |
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| 摘 要： | 目的构建针对Cav2.2e37a基因的RNAi慢病毒载体,为抑制Cav2.2e37a基因表达治疗神经痛的实验研究打下基础。方法针对已经筛选确定的Cav2.2e37a基因RNAi有效靶序列,构建pLL3.7-Cav2.2e37a干扰质粒,测序鉴定。pRsv-REV,pMDlg-pRRE,pMD2G,pLL3.7-Cav2.2e37a共转染293T细胞,Real-time PCR测定病毒转导滴度。结果成功构建Cav2.2e37a shRNA的慢病毒载体LVshCav2.2e37a。浓缩病毒悬液的滴度为1×10^9Tu/ml。结论成功构建了Cav2.2e37a基因的RNAi慢病毒载体。
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| 关 键 词： | RNA干扰 Car2．2 e37a 慢病毒属 |
Construction and identification of RNA interference lentivirus vector of Cav2.2 e37a gene |
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| WANG Wei,FENG Ze-guo,CHEN Na,MA Tao,ZHOU Jian-ping.Construction and identification of RNA interference lentivirus vector of Cav2.2 e37a gene[J].Academic Journal of Pla Postgraduate Medical School,2010,31(10):1016-1018. |
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| Authors: | WANG Wei FENG Ze-guo CHEN Na MA Tao ZHOU Jian-ping |
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| Affiliation: | 1.Department of Anaesthesia, Chinese PLA General Hospital, Beijing 100853, China; 2.Department of Anesthesia, General Hospital of PLA Second Artillery Forces, Beijing 100088, China; 3.Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China) |
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| Abstract: | Objective To construct the RNA interference（RNAi） lentivirus vector of Cav2.2 e37a gene in order to inhibit the expression of Cav2.2 e37a gene. Methods An interfere plasmid vector, pLL3.7-Cav2.2e37a, was constructed and confirmed by sequencing according to the effective sequence of siRNA targeting Cav2.2 e37a gene identified in our previous study. 293 T cells were co-transfected with pRsv-REV, pMDlg-pRRE, pMD2G, and pLL3.7-Cav2.2e37a. Concentrated virus titer was detected by realtime PCR. Results The RNAi vector of Cav2.2 e37a gene（pLL3.7-Cav2.2e37a） producing Cav2.2 e37a shRNA was constructed. The concentrated titer of virus suspension was 1×10^9Tu/ml. Conclusion RNAi lentivirus vector of Cav2.2 e37a gene can be constructed. |
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| Keywords: | RNA Interference Cav2 2 e37a Lentivirus |
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